| 孙建立,马志强.miR-187通过靶向胰岛素生长因子 -1受体基因对人乳腺癌细胞增殖和凋亡的影响[J].安徽医药,2021,25(5):882-886. |
| miR-187通过靶向胰岛素生长因子 -1受体基因对人乳腺癌细胞增殖和凋亡的影响 |
| miR-187 regulates proliferation and apoptosis of breast cancer cells by targeting IGF-1R gene |
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| DOI：10.3969/j.issn.1009-6469.2021.05.009 |
| 中文关键词: 乳腺肿瘤 受体， IGF1型 miR-187 乳腺癌细胞 增殖 凋亡 |
| 英文关键词: Breast neoplasms Receptor, IGF Type 1 miR-187 Breast cancer cells Proliferation Apoptosis |
| 基金项目:开封市科技发展计划项目( 1703010) |
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| 中文摘要: |
| 目的研究 miR-187通过靶向胰岛素生长因子 -1受体( IGF-1R)基因对人乳腺癌 MDA-MB-231细胞增殖和凋亡的影响。方法采用实时荧光定量 PCR(qRT-PCR)检测人正常乳腺上皮细胞 HBL-100和不同乳腺癌细胞中 miR-187的表达水平。在 MDA-MB-231细胞中分别转染 miR-187 mimics(miR-187组)或阴性对照 mimics-NC(miR-NC组)以未转染的 MDA-MB-231细胞为空白对照(空白对照组)。采用 qRT-PCR和蛋白免疫印迹( Western blotting)法分别检测转染后，细胞中 miR-187和 IGF1R蛋白表达水平，噻唑蓝( MTT)法和流式细胞术分别检测细胞增殖和凋亡情况。 Western blot检测与增殖和凋亡相关蛋白表达水平。采用双荧光素酶报告基因实验验证 miR-187与 IGF-1R靶向调控关系。结果与人正常乳腺上皮细胞 HBL-100(1.00±0.10)相比，乳腺癌细胞中 miR-187表达水平明显降低( P<0.05)在 MDA-MB-231细胞中表达水平最低( 0.11±0.01)。与 miR-NC组和空白对照组相比， miR-187组的 miR-187表达水平明显上调，［( 0.98±0.09)、(1.00±0.11)比( 3.18±0.33)P<0.05］， IGF-1R蛋白表达水平明显下调［( 0.28±0.03)、(0.27±0.03)比( 0.09±0.01)P<0.05］，细胞增殖活力 OD值［( 0.79±0.08)、(，0.82±0.08)比( 0.43±0.04)］和 PCNA表达水平［(0.39±0.04)(0.40±0.04)比( 0.20±002)］均显著降低( P<0.05)凋亡率［(5.02±1.13)%、.，(4.61±1.01)%比( 20.38±3.44)%］和 Cleaved Caspase-3达水平［( 0.09±0.02)、(0.08±0.02)比( 0.58±0.07］显著升高( P<0.05)。双荧光素酶报告基因实验显示 miR-187与 IGF-1R基因 3’非编码区( UTR)存在靶向关系。结论 miR-187对乳腺癌 MDA-MB表、)，231细胞具有增殖抑制和凋亡诱导作用，其作用机制与靶向抑制 IGF-1R基因表达有关。 |
| 英文摘要: |
| Objective To investigate the effect of miR-187 on the proliferation and apoptosis of human breast cancer MDA-MB-231 cells by targeting insulin growth factor-1 receptor (IGF-1R) gene.Methods Real-time quantitative PCR (qRT-PCR) was used to detect the expressions of miR-187 in human normal mammary epithelial cells HBL-100 and different breast cancer cells. We transfected miR 187 mimics (miR-187 group) or adopted mimics-NC as negative control (miR-NC group) in MDA-MB-231 cells, and adopted non-transfected MDA-MB-231 cells as blank controls (control group). The expression levels of miR-187 and IGF-1R protein in transfected cells were detected by qRT-PCR and Western blot. Cell proliferation and apoptosis were detected by MTT assay and flow cytometry, respectively. Western blot was used to detect the expression levels of proteins related to proliferation and apoptosis. The dual luciferase reporter gene assay was used to verify the regulatory relationship between miR-187 and IGF-1R.Results Compared with human normal mammary epithelial cells HBL-100(1.00±0.10), the expression level of miR-187 was significantly decreased in breast cancer cells (P< 0.05), and the lowest level was found in MDA-MB-231 cells(0.11±0.01). Compared with miR-NC group and control group, miR-187 expression level was significantly up-regulated in miR-187 group [(0.98±0.09),(1.00±0.11)vs.(3.18±0.33),P<0.05], IGF-1R protein expression level was significantly down-regulated [(0.28±0.03),(0.27±0.03)vs.(0.09±0.01),P<0.05], cell proliferation activity OD value [(0.79± 0.08),(0.82±0.08)vs.(0.43±0.04)] and PCNA expression level [(0.39±0.04),(0.40±0.04)vs.(0.20±0.02)] were significantly decreased (P< 0.05), and the apoptotic rate [(5.02±1.13)%,(4.61±1.01)% vs. (20.38±3.44)%] and the expression of Cleaved Caspase-3 [(0.09±0.02), (0.08±0.02)vs.(0.58±0.07)] were significantly increased (P<0.05). The dual luciferase reporter gene assay showed that miR-187 had a targeting relationship with the 3′ UTR of the IGF-1R gene.Conclusion miR-187 can inhibit proliferation and induce apoptosis of breast cancer MDA-MB-231 cells, and its mechanism is related to the targeted inhibition of IGF-1R gene expression. |
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